HUBUNGAN FILOGENETIK DUA POPULASI KATAK POHON Nyctimystes infrafrenatus infrafrenatus (TYLER 1971)

DI INDONESIA BERDASARKAN GEN 16S rRNA

ZOOGEOGRAPHY (BIO 562)

GRADUATE SCHOOL

IPB UNIVERSITY

Taufan Nurzaman Sulaeman

(G352180071)

Dr. Ir. Achmad Farajallah, MSi.

Dr. Amir Hamidy, MSc.

Sekolah Pascasarjana

Institut Pertanian Bogor

2020

Nyctimystes infrafrenatus (Gunther 1867)Katak Pohon Hijau Papua | White Lipped Treefrog| Australian Giant Treefrog

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Bibir bawah (infralabial) berwarna putih. Strip putih memanjang hingga ke bagian bawah tympanum

Dorsal berwarna hijau

Dorsal berwarna putih

SVL = 130 mm (Menzies 2006)

Jari kaki depan berselaput sebagianJari kaki belakang

Berselaput penuh

Litoria

Nyctimystes

Ranoidea

Pyron dan Wiens 2011Duellman et al. 2016)

Latar Belakang

3

Permasalahan Sistematik :Morfologi : Litoria Molekuler : Nyctimystes(Menzies 2006; Pyron dan Wiens 2011; Duellman et al. 2016)

Distribusi luas :Wilayah timur Indonesia, Papua Nuginiutara Australia, Kaledonia Baru dan Kepulauan Solomon(Iskandar dan Colijn 2000; Menzies 2006; Iskandar et al. 2017; Frost 2020)

Peta distribusi Nyctimystes infrafrenatus

Species Kriptik (?)Chalcorana chalconota (Inger et al. 2009)

Rhacophorus dugritei (Li et al. 2012)

Polypedates leucomystax(Kuraishi et al. 2013)

Leptobrachium hasseltii(Hamidy dan Matsui 2017) Microhyla complex (Atmadja et al. 2018).

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Kepulauan Maluku : Kei, Ambon, Obi, Buru, Seram, Talaud, Morotai, Halmahera

Papua :Papua, Papua Barat, Aru, Waigeo, Misol, Tanimbar

(Brongersma 1948; Boulenger 1912; Tyler 1978; Setiadi dan Hamidy 2006; Hamidy dan Mulyadi 2007; Koch et al. 2007; Karin et al. 2018; Nugroho et al. 2019).

Latar Belakang

2 Subspesies:Nyctimystes infrafrenatus infrafrenatusNyctimystes infrafrenatus militarius(Tyler 1971)

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Subspesies : Nyctimystes infrafrenatus infrafrenatus

Sinonim :

Calamites dolichopsisLitoria guttataHyla dolichopsi var. tennuigranulataHyla spengeli) (Loveridge 1948; Tyler 1971; Iskandar dan Colijn 2000; Menzies 2006).

SubspesiesNyctimystes infrafrenatus militarius

Sinonim

Hyla militaria Hyla dolichopsi var. pollicaris

Latar Belakang

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Latar Belakang Nyctimystes infrafrenatus infrafrenatus

2 populasi alopatrik - 2 subregion

Nyctimystes infrafrenatus infrafrenatusPopulasi Maluku

Wallacea

Nyctimystes infrafrenatus infrafrenatusPopulasi Papua

Australo-Papua

• Sejarah geologi pembentukan kawasan yang berbeda (Hall 1998; Hall 2002; Hall

2009; Hall 2013).

• Amfibi endemik terhadap benua atau unit biogeografi tunggal (Vences dan

Kohler 2008

• Distribusi amfibi melalui laut terbatas (Duellman dan Trueb 1986; Vences et al. 2003; Fonte et

al. 2019)

Molekuler ???:

Gen 16S rRNA(Vences et al. 2005)

Morfometrik:Perbedaan pada ukuran tungkai belakang betina(Prafiadi et al. 2016)

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Latar Belakang

Permasalahan SistematikDistribusi luasDua populasi pada dua

subregion biogeografi Perbedaan morfometrik*

Tujuan

• Menentukan status taksonomi N. i. infrafrenatus • Mengevaluasi hubungan kekerabatan antara populasi

N. i. infrafrenatus Papua dan Maluku

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Waktu dan Tempat• November 2019 – April 2020• Laboratorium Herpetologi, Lembaga Ilmu Penelitian Indonesia (LIPI),

Cibinong, Bogor.

Sampel Penelitian

• Sediaan sekuens gen 16S rRNA Nyctimystes infrafrenatus infrafrenatus Laboratorium Herpetologi, LIPI

Primer 16S rRNA :

16 H-1 (5’- CTCCGGTCTGAACTCAGATCACGTAGG -3’)

16 L-1 (5’- CTGACCGTGCAAAGGTAGCGTAATCACT –3’)

• Sediaan Sekuens 16S rRNA Nyctimystes infrafrenatus infrafrenatus dan outgrup yang tersedia di Genbank (https://www.ncbi.nlm.nih.gov/genbank/)

Metode Penelitian

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METODE

1010

Sekuens LIPI

Sekuens Genbank

ALIGNMENT

MUSCLEPerangkat lunak : MEGA X

Metode Penelitian

Pensejajaran Sekuens Rekonstruksi Pohon Filogenetik Visualisasi

Perangkat lunak : FigTree

Maximum Likelihood (ML)

GTR + G1000 replikasi

Perangkat lunak : MEGA X

Bayesian Inference (BI)

GTR + G10.000.000 MCMC Frekuensi diagnosis 100.000Burn-in 25 %Topologi sampel tiap 1000 generasi

Perangkat lunak : MrBayes 3.2.7a

Jarak genetik(p-distance)

Hasil Pensejajaran

419 pb

1111

Table 1. Sampel N.i. infrafrenatus dan outgroup yang digunakan pada rekonstruksi pohon filogenetik.

Metode Penelitian

METODE

1212

Table 1 lanjutan.....

Metode Penelitian

Gambar 2. Pohon filogenetik berdasarkan topologi Bayesian Inference N. i. infrafrenatusdan outgrup berdasarkan sekuens 16Sr RNA (419 pb). Angka pada lengan pohon menunjukkan besarnya bootstrap ML (MLBS) dan probabilitas posterior (BPP).

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Hasil dan Pembahasan

• Monofiletik• Membentuk 2 subclade

A

B

Gambar 1. A) Spesimen N. i. infrafrenatus dari Timika, Papuamemiliki “bibir putih” yang lebih tebal dari N. i. infrafrenatusdari Ternate, Maluku. Foto : Amir Hamidy (A) dan M. Iqbal Setiadi (B).

Gambar 2. Pohon filogenetik berdasarkan topologi Bayesian Inference N. i. infrafrenatusdan outgrup berdasarkan sekuens 16Sr RNA (419 pb). Angka pada lengan pohon menunjukkan besarnya bootstrap ML (MLBS) dan probabilitas posterior (BPP).

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Table 2. Jarak genetik (uncorrected p-distances) diantara populasi N. i. infrafrenatus dan spesies lain dari genus Nycimystes

Ranoidea bella sp. nov danRanoidea auae 2.00 %(McDonald et al. 2016)

• Jarak genetik 3% , indikasi pemisahan tingkat species (Fouquet et al. 2007)

• Leptobrachium hasseltii Bali dan Jawa 3-4%(Hamidy dan Matsui 2017)

Hasil dan Pembahasan

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Perubahan permukaan air laut dan garis pantai pada masa glasiasi maksimum terakhir (LGM) memunculkan dangkalan Sahul (Yokoyama et al. 2001)

20 mya 15 mya

10 mya 5 mya

(18,000 14C yr BP) (16,000 14C yr BP)

(12,000 14C yr BP) (9000 14C yr BP) (6000 14C yr BP)

P-distance :

1.3-1.5% ]

Vicariance memisahkan Papua dari Australia(Hantoro et al. 1995; Van der Kaars 1991; Voris 2000)

Hasil dan Pembahasan

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Perubahan permukaan air laut dan garis pantai pada masa glasiasi maksimum terakhir (LGM) memunculkan dangkalan Sahul (Yokoyama et al. 2001)

20 mya 15 mya

10 mya 5 mya

(18,000 14C yr BP) (16,000 14C yr BP)

(12,000 14C yr BP) (9000 14C yr BP) (6000 14C yr BP)

Vicariance memisahkan Papua dari Australia(Hantoro et al. 1995; Van der Kaars 1991; Voris 2000)

Hasil dan Pembahasan

Jembatan distribusi/pertukaran Spesies• hutan tropis

dataran rendah• padang rumput-semak• sistem sungai (Hantoro et al. 1995; Van der Kaars 1991; Voris 2000)

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Gambar Palaeogeografi Wallacea(Hall 2013)

20 mya 15 mya

10 mya 5 mya

Halmahera adalah Oceanic Islands(Hall 1998; Hall 2002; Hall 2009; Lohman et al. 2011; Hall 2013).

Mekanisme distribusi spesies pada

oceanic island :

overseas/overwater jump dispersal”

Katak :

Boophis dan Mantidactylus

Madagascar Comoros(Vences et al. 2003; Fonte et al. 2019).

Hasil dan Pembahasan

Tropicagama temporalis (Kadal)Kolonisasi Maluku dari Australia melalui Papua (Karin et al. 2020)

Simpulan

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• Populasi N. i. infrafrenatus di Indonesia merupakan Clademonofiletik dengan dua subclade

• Berdasarkan pemisahan subclade pada pohon filogenetik dan jarakgenetik diatas ambang batas spesies, populasi N. i. infrafrenatusMaluku merupakan spesies yang berbeda dari populasi Papua.

• Secara morfologi terdapat perbedaan tebal garis putih padainfralabial yang dapat menjadi salah satu karakter diagnostik untukpendeskripsian spesies lebih lanjut.

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